Introduction

Methods

Dissections

Fixing

Antibodies

Mounting

Microscopy

Fluorescence and brightfield microscopy
After mounting, we waited an appropriate amount of time for the slides to dry. All slides were initially looked at under a Zeiss Axiovert 200 fluorescence microscope. We viewed a total of 59 stained slides under the fluorescent microscope and chose selected slides to be imaged in more detail using an Olympus Fluoview 1000 confocal microscope.

Confocal microscopy
After locating the area of interest on each slide using regular fluorescence and brightfield microscopy we used the confocal imaging software. To optimize the image we adjusted the focus, offset, voltage, and gain. To generate the images below we used a Kalman filter to minimize background noise and a longer exposure time to obtain high quality images. We additionally took Z stacks of the slides in order to view a 3D perspective of the tissue.